1. Field of the Invention
The present invention relates to a laser scanning microscope for observing a specimen by emitting a laser beam onto the specimen marked with a plurality of fluorescent probes, scanning an observing plane of the specimen in two dimensions, and receiving a fluorescent light from the specimen; and in particular to a spectral data acquisition technique for such a laser scanning microscope.
2. Description of the Related Art
There is a laser scanning microscope which enables an observation by displaying in such as a monitor, through a process of generating an electric signal corresponding to a received fluorescent light back from a specimen by emitting a laser beam onto the specimen marked with a fluorescent probe and creating an image data corresponding to the fluorescent light back from the specimen based on the electric signal.
In order to observe a fluorescent light back from a specimen introduced by a fluorescent probe by using a laser scanning microscope, it is necessary to use a pump laser, photometric dichroic mirror and absorption filter, all of which match with an excitation wavelength and fluorescent peak wavelength of the fluorescent probe.
FIG. 1 is a block diagram showing a configuration of such conventional laser scanning microscope.
As shown by FIG. 1, depending on categories of fluorescent probes for marking a specimen, a plurality of following components, i.e., laser sources 87 through 89 for emitting pump lasers, photometric dichroic mirrors 91 and absorption filters 92, are furnished.
Let it first describe an operation of the laser scanning microscope shown by FIG. 1.
First, the laser sources 87 through 89 emit pump lasers which are focused and synthesized by way of condenser apparatuses 85 and synthesis mirror 86, and which are then emitted onto a specimen (plane) after being transmitted by way of a total reflection mirror 84, excitation dichroic mirror 83, deflection unit 82 and object lens 81.
The fluorescent light back from the specimen corresponding to the emission of pump laser transmits itself by way of the object lens 81 and the deflection unit 82 to reach at the excitation dichroic mirror 83.
The fluorescent light back from the specimen is reflected by the excitation dichroic mirror 83 and converted into electric signals at photoelectric conversion units 93, respectively, after going by way of the photometric dichroic mirror 91, which selects and disperses the fluorescent light from the fluorescent probe, and absorption filters 92 so that a display monitor (not shown herein) displays an image corresponding to the specimen (e.g., cell) as a subject of observation based on the electric signal.
And a selective switching of the combination between the above described photometric dichroic mirror 91 and absorption filter 92 enables a detection of fluorescent light from among the plurality of fluorescent probes.
In the meantime, a patent document noted below has disclosed a technique for automatically setting the optimum combination among an optimal pump laser, photometric dichroic mirror and absorption filter based on a pump laser equipped in a microscope system, spectral data of various filters, excitation wavelength data of fluorescent probes introduced to a test sample (i.e., specimen) and fluorescent wavelength data.
[Patent document 1] Japanese patent laid-open application publication No. 2000-39563: “Method and system configuration for adjusting equipment arrangement for confocal microscope”